Immunomodulatory Effects of Calcitriol through DNA Methylation Alteration of FOXP3 in the CD4+ T Cells of Mice
Vitamin D plays a variety of physiological functions, such as regulating mineral homeostasis. More recently, it has emerged as an immunomodulator player, affecting several types of immune cells, such as regulatory T (Treg) cells. It has been reported that vitamin D exerts some mediatory effects through an epigenetic mechanism. In this study, the impacts of calcitriol, the active form of vitamin D, on the methylation of the conserved non-coding sequence 2 (CNS2) region of the forkhead box P3 (Foxp3) gene promoter, were evaluated.
Fourteen C57BL/6 mice were recruited in this study and divided into two intervention and control groups. The CD4+ T cells were isolated from mice splenocytes. The expression of Foxp3, IL-10, and transforming growth factor-beta (TGF-β1) genes were relatively quantified by real-time PCR technique, and the DNA methylation percentage of every CpG site in the CNS2 region was measured individually by bisulfite-sequencing PCR.
Vitamin D Intervention significantly (p<0.05) could increase the expression of Foxp3, IL-10, and TGF-β1 gene in the CD4+ T cells of mice comparing with the control group. Meanwhile, methylation of the CNS2 region of Foxp3 promoter was significantly decreased in three of ten CpG sites in the vitamin D group compared to the control group.
The results of this study showed that vitamin D can engage the methylation process to induce Foxp3 gene expression and probably Treg cytokines profile. Further researches are needed to discover the precise epigenetic mechanisms by which vitamin D modulates the immune system.
2. Wei R, Christakos S. Mechanisms underlying the regulation of innate and adaptive immunity by vitamin D. Nutrients 2015;7(10):8251-60.
3. Penna G, Roncari A, Amuchastegui S, Daniel KC, Berti E, Colonna M, et al. Expression of the inhibitory receptor ILT3 on dendritic cells is dispensable for induction of CD4+ Foxp3+ regulatory T cells by 1, 25-dihydroxyvitamin D3. Blood 2005;106(10):3490-7.
4. Barragan M, Good M, Kolls JK. Regulation of dendritic cell function by vitamin D. Nutrients 2015;7(9):8127-51.
5. Vanherwegen A-S, Eelen G, Ferreira GB, Ghesquière B, Cook DP, Nikolic T, et al. Vitamin D controls the capacity of human dendritic cells to induce functional regulatory T cells by regulation of glucose metabolism. J Steroid Biochem Mol Biol 2019;187:134-45.
6. Pike JW, Meyer MB, Benkusky NA, Lee SM, John HS, Carlson A, et al. Genomic determinants of vitamin D-regulated gene expression. Vitam Horm 100: Elsevier; 2016. p. 21-44.
7. Carlberg C, Molnár F. Vitamin D receptor signaling and its therapeutic implications: Genome-wide and structural view. Can J Physiol Pharmacol 2015;93(5):311-8.
8. Heikkinen S, Väisänen S, Pehkonen P, Seuter S, Benes V, Carlberg C. Nuclear hormone 1α, 25-dihydroxyvitamin D3 elicits a genome-wide shift in the locations of VDR chromatin occupancy. Nucleic Acids Res 2011;39(21):9181-93.
9. Pereira F, Barbáchano A, Singh PK, Campbell MJ, Muñoz A, Larriba MJ. Vitamin D has wide regulatory effects on histone demethylase genes. Cell Cycle 2012;11(6):1081-9.
10. Ambrosi C, Manzo M, Baubec T. Dynamics and context-dependent roles of DNA methylation. J Mol Biol 2017;429(10):1459-75.
11. Bestor TH, Edwards JR, Boulard M. Notes on the role of dynamic DNA methylation in mammalian development. Proc Natl Acad Sci 2015;112(22):6796-9.
12. Sassi F, Tamone C, D’Amelio P. Vitamin D: nutrient, hormone, and immunomodulator. Nutrients 2018;10(11):1656.
13. Kang SW, Kim SH, Lee N, Lee W-W, Hwang K-A, Shin MS, et al. 1, 25-Dihyroxyvitamin D3 promotes FOXP3 expression via binding to vitamin D response elements in its conserved noncoding sequence region. J Immunol 2012;188(11):5276-82.
14. Zheng Y, Josefowicz S, Chaudhry A, Peng XP, Forbush K, Rudensky AY. Role of conserved non-coding DNA elements in the Foxp3 gene in regulatory T-cell fate. Nature 2010;463(7282):808-12.
15. Noori-Zadeh A, Mesbah-Namin SA, Saboor-Yaraghi AA. Epigenetic and gene expression alterations of FOXP3 in the T cells of EAE mouse model of multiple sclerosis. J Neurol Sci 2017;375:203-8.
16. Jiang M, Zhang Y, Fei J, Chang X, Fan W, Qian X, et al. Rapid quantification of DNA methylation by measuring relative peak heights in direct bisulfite-PCR sequencing traces. Lab Invest 2010;90(2):282-90.
17. Tsunoda T, Takagi T.Estimating transcription factorbindability on DNA. Bioinformatics 1999;15(7-8):622-30.
18. Tao R, De Zoeten EF, Özkaynak E, Chen C, Wang L, Porrett PM, et al. Deacetylase inhibition promotes the generation and function of regulatory T cells. Nat Med 2007;13(11):1299-307.
19. Reilly CM, Thomas M, Gogal Jr R, Olgun S, Santo A, Sodhi R, et al. The histone deacetylase inhibitor trichostatin A upregulates regulatory T cells and modulates autoimmunity in NZB/W F1 mice. J Autoimmun 2008;31(2):123-30.
20. Saouaf SJ, Li B, Zhang G, Shen Y, Furuuchi N, Hancock WW, et al. Deacetylase inhibition increases regulatory T cell function and decreases incidence and severity of collagen-induced arthritis. Exp Mol Pathol 2009;87(2):99-104.
21. Spanier JA, Nashold FE, Mayne CG, Nelson CD, Hayes CE. Vitamin D and estrogen synergy in Vdr-expressing CD4+ T cells is essential to induce Helios+ FoxP3+ T cells and prevent autoimmune demyelinating disease. J Neuroimmunol 2015;286:48-58.
22. Wang Y, Su MA, Wan YY. An essential role of the transcription factor GATA-3 for the function of regulatory T cells. Immunity 2011;35(3):337-48.
23. Fu W, Ergun A, Lu T, Hill JA, Haxhinasto S, Fassett MS, et al. A multiply redundant genetic switch'locks in'the transcriptional signature of regulatory T cells. Nat Immunol 2012;13(10):972.
24. Ouaked N, Mantel P-Y, Bassin C, Burgler S, Siegmund K, Akdis CA, et al. Regulation of the foxp3 gene by the Th1 cytokines: the role of IL-27-induced STAT1. J Immunol 2009;182(2):1041-9.
25. Laurence A, Amarnath S, Mariotti J, Kim YC, Foley J, Eckhaus M, et al. STAT3 transcription factor promotes instability of nTreg cells and limits generation of iTreg cells during acute murine graft-versus-host disease. Immunity 2012;37(2):209-22.
26. Bansal AS, Henriquez F, Sumar N, Patel S. T helper cell subsets in arthritis and the benefits of immunomodulation by 1, 25 (OH) 2 vitamin D. Rheumatol Int 2012;32(4):845-52.
27. Lam E, Choi SH, Pareek TK, Kim B-G, Letterio JJ. Cyclin-dependent kinase 5 represses Foxp3 gene expression and Treg development through specific phosphorylation of Stat3 at Serine 727. Mol Immunol 2015;67(2):317-24.
28. Skrobot A, Demkow U, Wachowska M. Immunomodulatory role of vitamin D: a review. Current Trends in Immunity and Respiratory Infections: Springer; 2018. p. 13-23.
29. Urry Z, Chambers ES, Xystrakis E, Dimeloe S, Richards DF, Gabryšová L, et al. The role of 1α, 25-dihydroxyvitamin D 3 and cytokines in the promotion of distinct Foxp3+ and IL-10+ CD 4+ T cells. Eur J Immunol 2012;42(10):2697-708.
|Issue||Vol 19 No 5 (2020)|
|Calcitriol FOXP3 gene Methylation Regulatory T-lymphocytes Vitamin D|
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